FM4-64 and pfa fixing

[Jean-Pierre CLAMME]

Hi,

I have used FM4-64 to follow endocytosis in live cells but I was wondering if
anyone knows what would be the effect of fixation with just pfa (no
permeabilization) on the FM4-64 labeling. I know invitrogen has a fixable
version of FM1-43 (that I can’t use because of the color of my other dyes),
but what about FM4-64?

Thank you

[Aryeh Weiss]

The Zeiss Axiovert 100 series scopes use a rack and pinion arrangement to move
the turret in the Z-direction. One two of our scopes the rack (ie the toothed
rod that meshes with the axle which the focus knob rotates) comes loose and
fails to mesh properly with the pinion, so that the turret falls and cannot be
raised. No matter how hard I tighten it when I reseat it, it slips out of
position. This appears to have happened to a second Axiovert 100 that we have.

My question is: have other users on this list had this problem, and if so, how
was it resolved? I reported on this in 2001, and it worked for some years, but
now the problem has worsened and my attempts to tighten the rack do not work for
more than a few weeks.

BTW -- the repair procedure I described in 2001 assumes that one does not want
to move the scope  (for example, to preserve alignment). If one is willing the
move the scope,  then the rack can be accessed from the bottom without removing
the tube lens.

--aryeh
--
Aryeh Weiss
School of Engineering
Bar Ilan University
Ramat Gan 52900 Israel

Ph:  972-3-5317638
FAX: [phone]

[Sylvie Le Guyader-2]

Hi jp

There is also a fixable FM4-64. If I remember well PFA kills the
fluorescence. I have had a bit of trouble with fixable FM4-64 having a
rather broad excitation/emission and bleeding into other channels. I don’t
know if other people encountered the same problem.

Med vänlig hälsning / Best regards
 
Sylvie
 
@@@@@@@@@@@@@@@@@@@@@@@@
Sylvie Le Guyader
Dept of Biosciences and Nutrition
Karolinska Institutet
Novum
14157 Huddinge
Sweden
[phone]

> -----Original Message-----
> From: Confocal Microscopy List
> [mailto:[hidden email]] On Behalf Of jpclamme
> Sent: 13 January 2009 02:22
> To: [hidden email]
> Subject: FM4-64 and pfa fixing
>
> Hi,
>
> I have used FM4-64 to follow endocytosis in live cells but I was wondering

if
> anyone knows what would be the effect of fixation with just pfa (no
> permeabilization) on the FM4-64 labeling. I know invitrogen has a fixable
> version of FM1-43 (that I can’t use because of the color of my other
dyes),
> but what about FM4-64?
>
> Thank you

[Glen MacDonald-2]

Hello,
Try the fixable FM4-64, F34653.   We have had good results with it.    
the standard 4-64 is not retained after PFA, just like its cousin, 1-43

Regards,
Glen

Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Box 357923
University of Washington
Seattle, WA 98195-7923  USA
[phone]
[hidden email]

******************************************************************************
The box said "Requires WindowsXP or better", so I bought a Macintosh.
******************************************************************************


On Jan 12, 2009, at 5:22 PM, jpclamme wrote:

> Hi,
>
> I have used FM4-64 to follow endocytosis in live cells but I was  
> wondering if
> anyone knows what would be the effect of fixation with just pfa (no
> permeabilization) on the FM4-64 labeling. I know invitrogen has a  
> fixable
> version of FM1-43 (that I can’t use because of the color of my other  
> dyes),
> but what about FM4-64?
>
> Thank you

[Ian Dobbie]

Aryeh Weiss <[hidden email]> writes:

> The Zeiss Axiovert 100 series scopes use a rack and pinion arrangement
> to move the turret in the Z-direction. One two of our scopes the rack
> (ie the toothed rod that meshes with the axle which the focus knob
> rotates) comes loose and fails to mesh properly with the pinion, so
> that the turret falls and cannot be raised. No matter how hard I
> tighten it when I reseat it, it slips out of position. This appears to
> have happened to a second Axiovert 100 that we have.
>
> My question is: have other users on this list had this problem, and if
> so, how was it resolved? I reported on this in 2001, and it worked for
> some years, but now the problem has worsened and my attempts to
> tighten the rack do not work for more than a few weeks.

I have seen this several years ago. The situation was worsened by the
rack and pinion then not meshing properly and shaving falling off the
rack so that the microscope was not able to focus properly at some
positions even the the system properly tightened again. We had the
scope serviced by Zeiss who replaced the mechanism. I'm not sure if
they replaced both parts or just the rack.

Ian

[Don Elsmore]

Hello Ian,
 
Seen this problem a lot with 100 series.
You need to replace the brass rack as the teeth are usually damaged which causes this problem. These were available from Ziess.

Regards
 

Donald Elsmore

President/Field Service Engineer

 

Aventeck LLC

10112 Owen Brown Road,

Columbia, MD 21044

Cell: 443-745-2087

Fax: [phone]

www.aventeck.com

 

> Date: Fri, 27 Feb 2009 12:09:32 +0000
> From: [hidden email]
> Subject: Re: problem with Zeiss Axiovert 100 series scopes
> To: [hidden email]
>
> Aryeh Weiss <[hidden email]> writes:
>
> > The Zeiss Axiovert 100 series scopes use a rack and pinion arrangement
> > to move the turret in the Z-direction. One two of our scopes the rack
> > (ie the toothed rod that meshes with the axle which the focus knob
> > rotates) comes loose and fails to mesh properly with the pinion, so
> > that the turret falls and cannot be raised. No matter how hard I
> > tighten it when I reseat it, it slips out of position. This appears to
> > have happened to a second Axiovert 100 that we have.
> >
> > My question is: have other users on this list had this problem, and if
> > so, how was it resolved? I reported on this in 2001, and it worked for
> > some years, but now the problem has worsened and my attempts to
> > tighten the rack do not work for more than a few weeks.
>
> I have seen this several years ago. The situation was worsened by the
> rack and pinion then not meshing properly and shaving falling off the
> rack so that the microscope was not able to focus properly at some
> positions even the the system properly tightened again. We had the
> scope serviced by Zeiss who replaced the mechanism. I'm not sure if
> they replaced both parts or just the rack.
>
> Ian

---

MSN NZ Travel Get inspired - dream, research, plan and book your next holiday online.

[Aryeh Weiss]

I actually replaced the rack, but the problem returned a few weeks later.
I found that no matter how hard I tightened the rack, it would slide back
away from the pinion with time (and not very much time). Apparently, the static
friction against the wall of the scope is insufficient against the shear force.

I thought of making one with a tighter tolerance on the through holes for the
screws which hold it.  Our machinist said that this will not work, because
there has to be some play in  those holes in order to position the rack
correctly. I thought that a drop of superglue might help, but I am reluctant
to do something that might be irreversible. This problem has rendered the scope
useless, and is driving me crazy, because I dont know how it ever worked,
and why Zeiss made something like this.

--aryeh

Don Elsmore wrote:

> Hello Ian,
>  
> Seen this problem a lot with 100 series.
> You need to replace the brass rack as the teeth are usually
> damaged which causes this problem. These were available from Ziess.
>
> Regards
>  
> Donald Elsmore
> President/Field Service Engineer
>  
> Aventeck LLC
> 10112 Owen Brown Road,
> Columbia, MD 21044
> Cell: 443-745-2087
> Fax: [phone]
> www.aventeck.com
>
>
>
>  
>  > Date: Fri, 27 Feb 2009 12:09:32 +0000
>  > From: [hidden email]
>  > Subject: Re: problem with Zeiss Axiovert 100 series scopes
>  > To: [hidden email]
>  >
>  > Aryeh Weiss <[hidden email]> writes:
>  >
>  > > The Zeiss Axiovert 100 series scopes use a rack and pinion arrangement
>  > > to move the turret in the Z-direction. One two of our scopes the rack
>  > > (ie the toothed rod that meshes with the axle which the focus knob
>  > > rotates) comes loose and fails to mesh properly with the pinion, so
>  > > that the turret falls and cannot be raised. No matter how hard I
>  > > tighten it when I reseat it, it slips out of position. This appears to
>  > > have happened to a second Axiovert 100 that we have.
>  > >
>  > > My question is: have other users on this list had this problem, and if
>  > > so, how was it resolved? I reported on this in 2001, and it worked for
>  > > some years, but now the problem has worsened and my attempts to
>  > > tighten the rack do not work for more than a few weeks.
>  >
>  > I have seen this several years ago. The situation was worsened by the
>  > rack and pinion then not meshing properly and shaving falling off the
>  > rack so that the microscope was not able to focus properly at some
>  > positions even the the system properly tightened again. We had the
>  > scope serviced by Zeiss who replaced the mechanism. I'm not sure if
>  > they replaced both parts or just the rack.
>  >
>  > Ian
>
> ------------------------------------------------------------------------
> MSN NZ Travel Get inspired - dream, research, plan and book your next
> holiday online. <http://travel.msn.co.nz/>

--
Aryeh Weiss
School of Engineering
Bar Ilan University
Ramat Gan 52900 Israel

Ph:  972-3-5317638
FAX: [phone]

Israel Society for Microscopy 2009 meeting website:
http://materials.technion.ac.il/ism/ISM2009.html

[Keith Morris]

Yep the focus on our Axiovert 100 S attached to our Bio-Rad Radiance
confocal at UCL was always failing [once or twice a year]. This Axiovert had
a Bio-Rad motorised Z motor bolted onto the manual focus. We had years of
grief with this as the Bio-Rad maintenance contract only extended to the Z
motor, and somewhere internally was an invisible line where the problem
became Zeiss not Bio-Rad. Occasionally we had to get both a Bio-Rad confocal
and a Zeiss microscope Engineer together at the same time as both would
blame the other suppliers component for the slipping Z motor control. As we
had to pay by the hour for the Zeiss engineer and wait weeks to get him [as
we had no maintenance contract on that bit] it was a real pain. This carried
on when Bio-Rad became Zeiss MicroScience [the irony being lost on them]. We
ended up with new Zeiss manual focus bits and an expensive new Bio-Rad z
motor, after which things were fine again [I left UCL in 2007]. Total cost
to us about £2,000 and a few weeks confocal downtime over two years,
excluding the £5k+ Bio-Rad z motor that was covered under the confocal
maintenance contract. Much of the problem seemed to be the way the manual
coarse and manual fine focus interacted with each other [from memory:
slipping via some sort of friction plate between the two]. Hopefully Zeiss
can still provide the required spare parts for your manual focus; it is an
elderly microscope now.

In comparison our Zeiss Axiovert 100 Ms with Zeiss internal motorised focus
and motorised diochroic slider control have worked totally reliably over the
years. Moral: get all the kit from the same manufacturer so one can't blame
the other and buy a fully motorised microscope [motors are cheaper than
over-engineered machined parts these days, plus it's far more flexible with
PC control]. We had other problems with the Axiovert 100 M though, mainly
the objectives slamming into the stage the stage when rotating, which was
nice - cured by washers under the stage. The 100 replacement, the Axiovert
200 M [we use with our Zeiss 510 confocal] seems a lot lot better, although
these are fully motorised. We did wonder if the problems we experienced with
our Axiovert 100s were a result of the fall of the Berlin wall and
re-integration of Zeiss Jena into Carl Zeiss.

Keith

---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford  OX3 7BN,
United Kingdom.

Telephone:  [phone]
Email:  [hidden email]
Web-pages: http://www.well.ox.ac.uk/cytogenetics/
 

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Aryeh Weiss
Sent: 28 February 2009 17:41
To: [hidden email]
Subject: Re: problem with Zeiss Axiovert 100 series scopes

I actually replaced the rack, but the problem returned a few weeks later.
I found that no matter how hard I tightened the rack, it would slide back
away from the pinion with time (and not very much time). Apparently, the
static
friction against the wall of the scope is insufficient against the shear
force.

I thought of making one with a tighter tolerance on the through holes for
the
screws which hold it.  Our machinist said that this will not work, because
there has to be some play in  those holes in order to position the rack
correctly. I thought that a drop of superglue might help, but I am reluctant
to do something that might be irreversible. This problem has rendered the
scope
useless, and is driving me crazy, because I dont know how it ever worked,
and why Zeiss made something like this.

--aryeh

Don Elsmore wrote:

> Hello Ian,
>  
> Seen this problem a lot with 100 series.
> You need to replace the brass rack as the teeth are usually
> damaged which causes this problem. These were available from Ziess.
>
> Regards
>  
> Donald Elsmore
> President/Field Service Engineer
>  
> Aventeck LLC
> 10112 Owen Brown Road,
> Columbia, MD 21044
> Cell: 443-745-2087
> Fax: [phone]
> www.aventeck.com
>
>
>
>  
>  > Date: Fri, 27 Feb 2009 12:09:32 +0000
>  > From: [hidden email]
>  > Subject: Re: problem with Zeiss Axiovert 100 series scopes
>  > To: [hidden email]
>  >
>  > Aryeh Weiss <[hidden email]> writes:
>  >
>  > > The Zeiss Axiovert 100 series scopes use a rack and pinion

arrangement
>  > > to move the turret in the Z-direction. One two of our scopes the rack
>  > > (ie the toothed rod that meshes with the axle which the focus knob
>  > > rotates) comes loose and fails to mesh properly with the pinion, so
>  > > that the turret falls and cannot be raised. No matter how hard I
>  > > tighten it when I reseat it, it slips out of position. This appears
to
>  > > have happened to a second Axiovert 100 that we have.
>  > >
>  > > My question is: have other users on this list had this problem, and
if
>  > > so, how was it resolved? I reported on this in 2001, and it worked
for

>  > > some years, but now the problem has worsened and my attempts to
>  > > tighten the rack do not work for more than a few weeks.
>  >
>  > I have seen this several years ago. The situation was worsened by the
>  > rack and pinion then not meshing properly and shaving falling off the
>  > rack so that the microscope was not able to focus properly at some
>  > positions even the the system properly tightened again. We had the
>  > scope serviced by Zeiss who replaced the mechanism. I'm not sure if
>  > they replaced both parts or just the rack.
>  >
>  > Ian
>
> ------------------------------------------------------------------------
> MSN NZ Travel Get inspired - dream, research, plan and book your next
> holiday online. <http://travel.msn.co.nz/>

--
Aryeh Weiss
School of Engineering
Bar Ilan University
Ramat Gan 52900 Israel

Ph:  972-3-5317638
FAX: [phone]

Israel Society for Microscopy 2009 meeting website:
http://materials.technion.ac.il/ism/ISM2009.html

[Aryeh Weiss]

Thank you for your detailed reply. I think that you are describing a different
problem. The fine focus on the Axiovert 100 series was a vernier friction  drive
  that felt great but was  absolutely unusable for any kind of Z reproducibility.
Biorad offered a replacement that  did away with the fine focus, and replaced
the drive on one side of the scope with a belt-drive reduction system that
directly drives the coarse focus. We have that upgrade, and it worked much
better. BTW -- Biorad had some really good mechanical design people.

My problem is inside the microscope, and is certainly a Zeiss issue -- not a
Biorad issue.

--aryeh

Keith Morris wrote:

> Yep the focus on our Axiovert 100 S attached to our Bio-Rad Radiance
> confocal at UCL was always failing [once or twice a year]. This Axiovert had
> a Bio-Rad motorised Z motor bolted onto the manual focus. We had years of
> grief with this as the Bio-Rad maintenance contract only extended to the Z
> motor, and somewhere internally was an invisible line where the problem
> became Zeiss not Bio-Rad. Occasionally we had to get both a Bio-Rad confocal
> and a Zeiss microscope Engineer together at the same time as both would
> blame the other suppliers component for the slipping Z motor control. As we
> had to pay by the hour for the Zeiss engineer and wait weeks to get him [as
> we had no maintenance contract on that bit] it was a real pain. This carried
> on when Bio-Rad became Zeiss MicroScience [the irony being lost on them]. We
> ended up with new Zeiss manual focus bits and an expensive new Bio-Rad z
> motor, after which things were fine again [I left UCL in 2007]. Total cost
> to us about £2,000 and a few weeks confocal downtime over two years,
> excluding the £5k+ Bio-Rad z motor that was covered under the confocal
> maintenance contract. Much of the problem seemed to be the way the manual
> coarse and manual fine focus interacted with each other [from memory:
> slipping via some sort of friction plate between the two]. Hopefully Zeiss
> can still provide the required spare parts for your manual focus; it is an
> elderly microscope now.
>
> In comparison our Zeiss Axiovert 100 Ms with Zeiss internal motorised focus
> and motorised diochroic slider control have worked totally reliably over the
> years. Moral: get all the kit from the same manufacturer so one can't blame
> the other and buy a fully motorised microscope [motors are cheaper than
> over-engineered machined parts these days, plus it's far more flexible with
> PC control]. We had other problems with the Axiovert 100 M though, mainly
> the objectives slamming into the stage the stage when rotating, which was
> nice - cured by washers under the stage. The 100 replacement, the Axiovert
> 200 M [we use with our Zeiss 510 confocal] seems a lot lot better, although
> these are fully motorised. We did wonder if the problems we experienced with
> our Axiovert 100s were a result of the fall of the Berlin wall and
> re-integration of Zeiss Jena into Carl Zeiss.
>
> Keith
>
> ---------------------------------------------------------------------------
> Dr Keith J. Morris,
> Molecular Cytogenetics and Microscopy Core,
> Laboratory 00/069 and 00/070,
> The Wellcome Trust Centre for Human Genetics,
> Roosevelt Drive,
> Oxford  OX3 7BN,
> United Kingdom.
>
> Telephone:  [phone]
> Email:  [hidden email]
> Web-pages: http://www.well.ox.ac.uk/cytogenetics/
>  
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]] On
> Behalf Of Aryeh Weiss
> Sent: 28 February 2009 17:41
> To: [hidden email]
> Subject: Re: problem with Zeiss Axiovert 100 series scopes
>
> I actually replaced the rack, but the problem returned a few weeks later.
> I found that no matter how hard I tightened the rack, it would slide back
> away from the pinion with time (and not very much time). Apparently, the
> static
> friction against the wall of the scope is insufficient against the shear
> force.
>
> I thought of making one with a tighter tolerance on the through holes for
> the
> screws which hold it.  Our machinist said that this will not work, because
> there has to be some play in  those holes in order to position the rack
> correctly. I thought that a drop of superglue might help, but I am reluctant
> to do something that might be irreversible. This problem has rendered the
> scope
> useless, and is driving me crazy, because I dont know how it ever worked,
> and why Zeiss made something like this.
>
> --aryeh
>
> Don Elsmore wrote:
>> Hello Ian,
>>  
>> Seen this problem a lot with 100 series.
>> You need to replace the brass rack as the teeth are usually
>> damaged which causes this problem. These were available from Ziess.
>>
>> Regards
>>  
>> Donald Elsmore
>> President/Field Service Engineer
>>  
>> Aventeck LLC
>> 10112 Owen Brown Road,
>> Columbia, MD 21044
>> Cell: 443-745-2087
>> Fax: [phone]
>> www.aventeck.com
>>
>>
>>
>>  
>>  > Date: Fri, 27 Feb 2009 12:09:32 +0000
>>  > From: [hidden email]
>>  > Subject: Re: problem with Zeiss Axiovert 100 series scopes
>>  > To: [hidden email]
>>  >
>>  > Aryeh Weiss <[hidden email]> writes:
>>  >
>>  > > The Zeiss Axiovert 100 series scopes use a rack and pinion
> arrangement
>>  > > to move the turret in the Z-direction. One two of our scopes the rack
>>  > > (ie the toothed rod that meshes with the axle which the focus knob
>>  > > rotates) comes loose and fails to mesh properly with the pinion, so
>>  > > that the turret falls and cannot be raised. No matter how hard I
>>  > > tighten it when I reseat it, it slips out of position. This appears
> to
>>  > > have happened to a second Axiovert 100 that we have.
>>  > >
>>  > > My question is: have other users on this list had this problem, and
> if
>>  > > so, how was it resolved? I reported on this in 2001, and it worked
> for
>>  > > some years, but now the problem has worsened and my attempts to
>>  > > tighten the rack do not work for more than a few weeks.
>>  >
>>  > I have seen this several years ago. The situation was worsened by the
>>  > rack and pinion then not meshing properly and shaving falling off the
>>  > rack so that the microscope was not able to focus properly at some
>>  > positions even the the system properly tightened again. We had the
>>  > scope serviced by Zeiss who replaced the mechanism. I'm not sure if
>>  > they replaced both parts or just the rack.
>>  >
>>  > Ian
>>
>> ------------------------------------------------------------------------
>> MSN NZ Travel Get inspired - dream, research, plan and book your next
>> holiday online. <http://travel.msn.co.nz/>
>
>

--
Aryeh Weiss
School of Engineering
Bar Ilan University
Ramat Gan 52900 Israel

Ph:  972-3-5317638
FAX: [phone]

Israel Society for Microscopy 2009 meeting website:
http://materials.technion.ac.il/ism/ISM2009.html

[Keith Morris]

I'm not sure that's totally the case [except when the Bio-Rad z motor did
fall apart]. Apparently the monster Bio-Rad confocal bolt on motor we had
for the Zeiss Axiovert-100 was never as successful as the one used for the
main microscope manufacturer Bio-Rad generally supplied [was it Olympus or
Nikon? can't remember] - well according to our Bio-Rad engineer anyway [and
apparently they sold very few for the Zeiss Axiovert 100 in the UK]. The
Prof insisted on Zeiss, as we got a job lot of Axiovert 100's at a good
price [and were generally quite happy them after a few initial problems were
sorted out]. I'm not a fan of bolt-on z motors though, and the Bio-Rad one
did little to help change my mind.

I do know we had problems with the Zeiss manual mechanism that actually
raised and lowered the nosepiece, well away from the fine/coarse focus
knobs, and these were the parts that were replaced [a rack or pinion bit?].
We still had the Zeiss fine manual focus fully working [well mostly] on the
left side, which naturally became a little stiff when the z motor fired up
for the z-stack [the only time the z motor was used].

Using our manual focus Nikon TE2000U inverted yesterday I did notice how
often you can hit the objective nosepiece into the stops trying to get focus
and such, and it makes a terrible noise when doing it. I guess that’s why
inverted manual focus's can take a hit, it's difficult to see where you are
going with the objective and I'm pretty sure a lot of users aren't as
careful with the focus as I am. Possibly that’s why I find internal
motorised focus's so reliable, when they hit the stop they just stop, with a
little 'please don't do that again' beep.

Keith
 

---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford  OX3 7BN,
United Kingdom.

Telephone:  [phone]
Email:  [hidden email]
Web-pages: http://www.well.ox.ac.uk/cytogenetics/
 
-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Aryeh Weiss
Sent: 03 March 2009 19:57
To: [hidden email]
Subject: Re: problem with Zeiss Axiovert 100 series scopes

Thank you for your detailed reply. I think that you are describing a
different
problem. The fine focus on the Axiovert 100 series was a vernier friction
drive
  that felt great but was  absolutely unusable for any kind of Z
reproducibility.
Biorad offered a replacement that  did away with the fine focus, and
replaced
the drive on one side of the scope with a belt-drive reduction system that
directly drives the coarse focus. We have that upgrade, and it worked much
better. BTW -- Biorad had some really good mechanical design people.

My problem is inside the microscope, and is certainly a Zeiss issue -- not a

Biorad issue.

--aryeh

Keith Morris wrote:
> Yep the focus on our Axiovert 100 S attached to our Bio-Rad Radiance
> confocal at UCL was always failing [once or twice a year]. This Axiovert
had
> a Bio-Rad motorised Z motor bolted onto the manual focus. We had years of
> grief with this as the Bio-Rad maintenance contract only extended to the Z
> motor, and somewhere internally was an invisible line where the problem
> became Zeiss not Bio-Rad. Occasionally we had to get both a Bio-Rad
confocal
> and a Zeiss microscope Engineer together at the same time as both would
> blame the other suppliers component for the slipping Z motor control. As
we
> had to pay by the hour for the Zeiss engineer and wait weeks to get him
[as
> we had no maintenance contract on that bit] it was a real pain. This
carried
> on when Bio-Rad became Zeiss MicroScience [the irony being lost on them].
We

> ended up with new Zeiss manual focus bits and an expensive new Bio-Rad z
> motor, after which things were fine again [I left UCL in 2007]. Total cost
> to us about £2,000 and a few weeks confocal downtime over two years,
> excluding the £5k+ Bio-Rad z motor that was covered under the confocal
> maintenance contract. Much of the problem seemed to be the way the manual
> coarse and manual fine focus interacted with each other [from memory:
> slipping via some sort of friction plate between the two]. Hopefully Zeiss
> can still provide the required spare parts for your manual focus; it is an
> elderly microscope now.
>
> In comparison our Zeiss Axiovert 100 Ms with Zeiss internal motorised

focus
> and motorised diochroic slider control have worked totally reliably over
the
> years. Moral: get all the kit from the same manufacturer so one can't
blame
> the other and buy a fully motorised microscope [motors are cheaper than
> over-engineered machined parts these days, plus it's far more flexible
with
> PC control]. We had other problems with the Axiovert 100 M though, mainly
> the objectives slamming into the stage the stage when rotating, which was
> nice - cured by washers under the stage. The 100 replacement, the Axiovert
> 200 M [we use with our Zeiss 510 confocal] seems a lot lot better,
although
> these are fully motorised. We did wonder if the problems we experienced
with
> our Axiovert 100s were a result of the fall of the Berlin wall and
> re-integration of Zeiss Jena into Carl Zeiss.
>
> Keith
>
>
---------------------------------------------------------------------------

> Dr Keith J. Morris,
> Molecular Cytogenetics and Microscopy Core,
> Laboratory 00/069 and 00/070,
> The Wellcome Trust Centre for Human Genetics,
> Roosevelt Drive,
> Oxford  OX3 7BN,
> United Kingdom.
>
> Telephone:  [phone]
> Email:  [hidden email]
> Web-pages: http://www.well.ox.ac.uk/cytogenetics/
>  
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]

On

> Behalf Of Aryeh Weiss
> Sent: 28 February 2009 17:41
> To: [hidden email]
> Subject: Re: problem with Zeiss Axiovert 100 series scopes
>
> I actually replaced the rack, but the problem returned a few weeks later.
> I found that no matter how hard I tightened the rack, it would slide back
> away from the pinion with time (and not very much time). Apparently, the
> static
> friction against the wall of the scope is insufficient against the shear
> force.
>
> I thought of making one with a tighter tolerance on the through holes for
> the
> screws which hold it.  Our machinist said that this will not work, because
> there has to be some play in  those holes in order to position the rack
> correctly. I thought that a drop of superglue might help, but I am

reluctant

> to do something that might be irreversible. This problem has rendered the
> scope
> useless, and is driving me crazy, because I dont know how it ever worked,
> and why Zeiss made something like this.
>
> --aryeh
>
> Don Elsmore wrote:
>> Hello Ian,
>>  
>> Seen this problem a lot with 100 series.
>> You need to replace the brass rack as the teeth are usually
>> damaged which causes this problem. These were available from Ziess.
>>
>> Regards
>>  
>> Donald Elsmore
>> President/Field Service Engineer
>>  
>> Aventeck LLC
>> 10112 Owen Brown Road,
>> Columbia, MD 21044
>> Cell: 443-745-2087
>> Fax: [phone]
>> www.aventeck.com
>>
>>
>>
>>  
>>  > Date: Fri, 27 Feb 2009 12:09:32 +0000
>>  > From: [hidden email]
>>  > Subject: Re: problem with Zeiss Axiovert 100 series scopes
>>  > To: [hidden email]
>>  >
>>  > Aryeh Weiss <[hidden email]> writes:
>>  >
>>  > > The Zeiss Axiovert 100 series scopes use a rack and pinion
> arrangement
>>  > > to move the turret in the Z-direction. One two of our scopes the

rack

>>  > > (ie the toothed rod that meshes with the axle which the focus knob
>>  > > rotates) comes loose and fails to mesh properly with the pinion, so
>>  > > that the turret falls and cannot be raised. No matter how hard I
>>  > > tighten it when I reseat it, it slips out of position. This appears
> to
>>  > > have happened to a second Axiovert 100 that we have.
>>  > >
>>  > > My question is: have other users on this list had this problem, and
> if
>>  > > so, how was it resolved? I reported on this in 2001, and it worked
> for
>>  > > some years, but now the problem has worsened and my attempts to
>>  > > tighten the rack do not work for more than a few weeks.
>>  >
>>  > I have seen this several years ago. The situation was worsened by the
>>  > rack and pinion then not meshing properly and shaving falling off the
>>  > rack so that the microscope was not able to focus properly at some
>>  > positions even the the system properly tightened again. We had the
>>  > scope serviced by Zeiss who replaced the mechanism. I'm not sure if
>>  > they replaced both parts or just the rack.
>>  >
>>  > Ian
>>
>> ------------------------------------------------------------------------
>> MSN NZ Travel Get inspired - dream, research, plan and book your next
>> holiday online. <http://travel.msn.co.nz/>
>
>

--
Aryeh Weiss
School of Engineering
Bar Ilan University
Ramat Gan 52900 Israel

Ph:  972-3-5317638
FAX: [phone]

Israel Society for Microscopy 2009 meeting website:
http://materials.technion.ac.il/ism/ISM2009.html